Acanthosis Nigricans ; diagnosis ; genetics ; pathology ; Adolescent ; Antigens, CD ; genetics ; Biomarkers ; blood ; DNA Mutational Analysis ; Donohue Syndrome ; diagnosis ; genetics ; pathology ; Humans ; Insulin Resistance ; genetics ; Male ; Mutation ; genetics ; Polymerase Chain Reaction ; Receptor, Insulin ; genetics ; Sequence Analysis, DNA

Objective To study the transcription level of exogenetic glial cell line-derived neurotrophic factor(GDNF)in rat bone marrow mesenchymal stem cells(BMSCs).Methods The rat BMSCs were isolated and cultured in vitro,and randomly divided into three groups:GDNF-transfected group,blank plasmid-transfected group and non-transfected group.GDNF-transfected group and blank plasmid-transfected group were infected by the recombinant adenovirus of GDNF gene and blank plasmid,respectively.Each of the groups was divided again into 4 subgroups according to the cultured time(3d,6d,9d and 12d)after infection.The transcription level of GDNF gene in BMSCs was tested by quantitative real-time PCR.Results Blank plasmid-transfected group and non-transfected group had no expression of GDNF mRNA,and the relative level of GDNF mRNA transcription was 0.00751?0.00067 in GDNF-transfected group during the following 3~12 days after transfection.Conclusion Gene transfection technoqie mediated by adenoviral vector can transfect GDNF gene into BMSCs with the expression of exogenetic GDNF mRNA.BMSCs transfected with GDNF gene will be used for further research on therapy for the disease of central nervous system.

Objective To investigate the feasibility of using ultrasound‐mediated destruction of microbubbles ( US+ MB) to enhance the transplantation of endothelial progenitor cells ( EPCs) to confer chronic allograft vasculopathy (CAV) .Methods Bone marrow derived mononuclear cells were isolated and induced in vitro . The abdominal aorta transplantation was performed . Four groups were divided:control group without treatment (group A) ,injection with saline (group B) ,injection with EPCs (group C) ,group D ( US+MB+EPCs) was injected with EPCs and US was applied to MB prior to the infusion . All rats were killed during 8 weeks after transplantation to enable histological examination;SDF‐1α expression was detected by immunohistochemistry ,the expression of SDF‐1αand TNF‐αin the grafted aortas were detected with RT‐PCR . Results When 8 weeks after EPCs transplantation ,there was a significant improvement in aortic intima of Group D compared with Group B and C ,respectively ( P <0 .05) . In addition ,treatment of Group D significantly increased the expression of SDF‐1αand reduced the expression of TNF‐αin the grafted aortas . Conclusions US‐mediated MB destruction prior to EPCs transplantation into the grafted aortas can improves the effectiveness of endothelial repair and delay the progress of CAV .

Aim To observe the effects of theaflavin on intracellular calcium concentration(i) level in rat ventricular myocytes and discuss the possible mechanisms.Methods The effects of theaflavin oni were investigated in rat ventricular myocytes.i was detected by laser confocal microscopy and represented by relative fluorescent intensity(FI-FI0)/FI0,%;FI0:control;FI:administration of drugs).Results ① Theaflavin(10,20,40 ?mol?L-1) had no effect on the i of ventricular myocytes in normal Tyrode′s solution.However,it reduced the i of ventricular myocytes in simulated ischemia solution in a concentration-dependent manner.② Pretreatment with Bay k8644(0.5 ?mol?L-1) mostly abolished the effects of theaflavin(20 ?mol?L-1) in simulated ischemia solution.③ Theaflavin(20 ?mol?L-1) markedly inhibited the low concentration of ryanodine-induced i increase in Ca2+-free Tyrode′s solution.④ When the propagating waves of elevated i(Ca2+ waves) were produced by increasing extracellular Ca2+ concentration from 1 mmol?L-1to 10 mmol?L-1,theaflavin(20 ?mol?L-1) could block the propagating waves of elevated i(Ca2+ waves),reduce the frequency and duration of propagating waves,and reduce i as well.Conclusion Theaflavin may reduce the i in isolated rat ventricular myocytes via inhibiting Ca2+ influx by voltage-dependent Ca2+ channel and alleviating Ca2+ release from sarcoplasmic reticulum(SR).

Objective To investigate the long-term prognosis an d recorery of children with juvenile rheumatoid arthritis(JRA).Methods The cases diagnosed JRA in our hospital over the period of 1988～1992 we re followed up for the conditions of disease,the deteriorated joints,the treatme nt and the living conditions of patients.Results Ninty-six ca ses were followed up(involving male 66 cases,female 29 cases,the mean age of on set 8.21?3.17 years )except for a case who died of lymphoma.There were 44 cases with systemic JRA ,2 cases of them died after 7-years onset and 23 cases developed severe destructive arthritis.There were 38 oligoarthritis cases,27 cases of them stiu had active disease during 10-year following-up and 9 cases were diagnosed sacroiliitis.There were 13 polyarthritis and 3 cases of them were RF positive, who had developed severe destructive arthritis.Conclusions The prognosis of juvenile rheumatoid arthritis is not desirable,especially in systemic JRA,whose prognostic factors are related to age of onset,the lasting of fever,the markers of phlegmasia activity and the condition of systemic involvement and treatment.Oligoarthritis about 30 % may develop into ankylosing spondylitis.The probability of destructive arthritis is hi gher in polyarthritis with more RF positive and poor prognosis.

leptin-induced VSMCs proliferation as well as expression of PCNA and OB-R at both mRNA and protein levels. The maximum effect was at 100 μmol/L(P<0.01).

Objective To evaluate clinical application of Jin Pujia GP HER2 probe kit in testing HER2 gene status of breast cancer through comparing it with PathVysion HER2 probe kit. Methods HER2 gene status were detected from 108 cases with invasive ductal breast cancer using GP and PathVysion HER2 probe kits by FISH. HER2 gene expression levels were measured by GP and PathVysion HER2 probe kits, and the sensitivity, the specificity and the accuracy of GP HER2 probe kit were evaluated. Results HER2 gene amplification positive rates detected by GP HER2 probe kit and PathVysion HER2 probe kit were 25.0%(27/108) and 26.9% (29/108), respectively. As compared with PathVysion HER2 probe kit, the sensitivity, the specificity and the accuracy of the GP HER2 kit were 89. 7% (26/29), 98.7% (78/79)and 96. 3% ( 104/108), respectively, whereas the PPV and NPV were 96. 3% (26/27) and 96. 3% (78/81), respectively. The GP HER2 probe kit had a sensitivity of 93.3% ( 14/15), a specificity of 100%(93/93) and an accuracy of 99. 1% (107/108) for detecting polysomy 17. Conclusion GP HER2 probe kit has high sensitivity and specificity for detecting HER2 gene status in breast cancer patients, and it has clinical application value.

Objective Autosomal dominant congenital cataract (ADCC) is a common heredit disease.Some known genes and mutated loci related to ADCC have been found.The present study provides other disease-causing genes in the ADCC family.This study was to identify the genetic defect in four generations of a Chinese family with autosomal dominant congenital membranous cataracts and demonstrate the functional analysis of a candidate gene in the family.MethodsThe family with hereditary cataract was recruited from the Tianjin Medical University Eye Center.The family history was collected and recorded.Clinical and ophthalmologic examinations were performed on 6 affected and 14 unaffected family members and periphery blood samples were collected from all of the subjects for genomic DNA preparation.The members were genotyped with microsatellite markers at loci associated with cataracts.Multiplex polymerase chain reaction (PCR) was carried out with microsatellite markers near to candidated loci related to congenital cataracts.PCR products from each DNA sample were separated on a polyarcylamide gel and analyzed.Exclusion analysis was performed by allele sharing analysis and gene sequencing.This trail was approved by the Human Research Ethics Committee of this hospital.The oral informed consent was obtained from all of the subjects before the initiation of this trial.ResultsThe hereditary characteristic of this family was in accordance with the autosomal dominant inheritance with a gene penetrance 100%.Affected members of the family were diagnosed with membranous cataracts without other ocular symptom.The disease-causing gene locus were mapped to 22q11.2-q12.1 at a size of about 2.4 Mbp.The multiple-sequence alignments of complete coding region and splice site of CRYBB1,CRYBB2,CRYBB3,CRYBA4 were obtained but no mutation was found in this study.CRYBB1,CRYBB2,CRYBB3,CRYBA4 were screened by directly sequencing.ConclusionAll known ADCC loci have been excluded from the family.Further study should be carried out to screen other relevant genes or loci in patients with ADCC.The pathogenic gene in the family should be identified through extensive scanning of genes,and a new disease-causing gene may exist in this family.

Objective: To investigate the effect of sevoflurane anesthesia on hepatic ischemia-reperfusion injury in patients undergoing partial hepatectomy. Methods: A total of 86 patients undergoing partial hepatectomy were divided into sevoflurane group and isoflurane group according to random number table with 43 patients in each group. Both groups received continuous epidural block combined with general anesthesia, sevoflurane group took sevoflurane combined with remifentanil to maintain anesthesia, while isoflurane group took isoflurane remifentanil to maintain anesthesia. The mean arterial pressure (MAP) and heart rate(HR) of different moments, awakening (open-eye time, extubation time, listening to instruction time, orientation recovery time, time of Aldrete score up to 9 minutes), the serum levels of tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), aspartate aminotransferase (AST), alanine aminotransferase (ALT), glutamate dehydrogenase (GLDH), alpha glutathione S transferase (α-GST) levels at different moments were compared between the two groups. Results: The MAP and HR at T₁, T₂, T₃ and T₄ in both groups were significantly lower than those at T₀ (P<0.05). The HR at T₁ and T₄ in sevoflurane group were significantly higher than those in isoflurane group (P<0.05). The open-eye time, extubation time, listening to instruction time, orientation recovery time in sevoflurane group were significantly shorter than the isoflurane group (P<0.05). The levels of serum TNF-α, IL-1, AST, ALT, GLDH and α-GST in the two groups at T₂ and T₃ were significantly higher than those at T₀ (P<0.05), and began to fall back to T₄. The levels of serum TNF-α, IL-1, AST, ALT, GLDH and α-GST in the sevoflurane group at T₂ and T₃ were significantly lower than those in the isoflurane group (P<0.05). Conclusions The hepatic hemodynamics in patients undergoing partial hepatectomy with sevoflurane is more stable better than isoflurane during operation, which has faster postoperative recovery and relieved hepatic ischemia-reperfusion injury.

Objective To evaluate the change of left ventricular diastolic function and investigate the relation between left ventricular diastolic function and disease activity in rheumatoid arthritis (RA) without clinical manifestations of heart diseases. Methods Seventy consecutive active RA in-patients without clinical manifestations of heart disease were enrolled, while the control group was recruited from outpatient health physical check-up center and consisted of 60 age- and sex-matched healthy subjects. Cardiac related parame-ters were determined by echocardiography and the correlation between left ventricular diastolic function and the disease activity indexes were evaluated. Chi-square test, t test, Pearson or Spearman′s correlation test and Stepwise backward linear regression analysis were used for statistical analysis. Results RA patients had lower mitral inflow E/A ratio (1.2±0.4, 1.5±0.4, P<0.01), higher E/Em ratio (9.6±3.7, 7.8±2.0, P<0.01), longer isovolumetric relaxation time(IVRT)[(64±16) ms,(58±16) ms, P<0.05] than control group. Whilst, RA patients had higher pulmonary venous inflow A wave velocity-time integral (ArVTI) and A wave duration (DAr)[3.2±0.7,(2.8±0.6) cm; 117±11,(102±9) ms, P<0.05]. Moreover, the E/Em was positively corre-lated with C-reactive protein(CRP)(r=0.581, P<0.01), DAS28(r=0.456, P<0.01). Anti-CCP level was also associated with Em and early diastolic pulmonary venous inflow peak velocity(PVD)(r=-0.359, P<0.05;r=-0.305, P<0.05). In addition, multivariate analysis also revealed that there was linear regression relation-ship between E/Em and CRP, DAS28(t=3.266, P=0.002; t=2.949, P=0.005). Conclusion The study has revealed that left ventricular diastolic function is impaired in RA patients and the left ventricular diastolic function parameters is associated with the disease activity indexes. These results suggest that the decline of left ventricular diastolic function is associated with the inflammation activity in RA patients without clinical manifestations of heart disease.